Organ-Specific Protein Detection via High-Abundance Removal
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Solution Overview
Problem
Current diagnostic methods for detecting diseases, particularly cancers, face challenges in resolving large numbers of proteins in blood samples due to high abundance proteins distorting gel patterns, making it difficult to identify low abundance proteins and achieve reproducibility, especially in techniques like 2-DE gel electrophoresis, which limits early disease detection and diagnostic assays.
Innovation Solution
A method involving generating signature sequences from transcripts of specific organs, identifying organ-specific proteins secreted into the blood by comparing these sequences to databases, and using detection reagents like antibodies or aptamers to measure protein levels in blood samples for diagnostic purposes, enabling the identification of organ-specific molecular blood fingerprints.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If 2-DE gel electrophoresis is used to analyze blood proteins, then protein separation is achieved, but high abundance proteins distort carrier ampholyte gradients and cause crowding in the gel matrix, resulting in irreproducibility
Solution Approach 1:
The patent extracts and removes high abundance proteins (such as albumin) from the blood sample before performing gel electrophoresis. This is achieved through pre-clearance steps using affinity columns or other separation methods that selectively remove these interfering proteins, thereby preventing them from distorting the gel matrix and carrier ampholyte gradients during electrophoresis.
Solution Approach 2:
The patent performs preliminary processing of the blood sample to remove high abundance proteins before the actual gel electrophoresis analysis. This preliminary action includes using affinity columns or other separation techniques to pre-clear the sample, ensuring that the subsequent gel analysis is not compromised by the presence of abundant proteins that would cause crowding and irreproducibility.
2Measurement precision
If 2-DE gel electrophoresis is used to resolve proteins, then protein separation is achieved, but low abundance proteins cannot be resolved from high abundance proteins due to high staining background and limited dynamic range
Solution Approach 1:
The patent extracts and removes high abundance proteins from the blood sample before gel electrophoresis. By using affinity columns or other separation methods to pre-clear the sample of proteins like albumin, the technique eliminates the high staining background that would otherwise mask low abundance proteins, thereby enabling their detection.
Solution Approach 2:
The patent performs preliminary sample processing to remove high abundance proteins before gel analysis. This pre-clearance step using affinity columns or other separation techniques ensures that the gel staining background is reduced, allowing low abundance proteins to be visualized and detected with sufficient sensitivity.
3Productivity
If conventional gel electrophoresis is used for diagnostic proteomics, then protein analysis is performed, but the complexity of the blood proteome and large dynamic range (10^10) make it difficult to identify useful diagnostic markers
Solution Approach 1:
The patent segments the blood proteome analysis into distinct functional steps: (1) pre-clearance to remove high abundance proteins, (2) gel electrophoresis for separation of remaining proteins, and (3) staining and detection. This segmentation simplifies the overall process by handling the complex proteome in manageable stages rather than attempting to analyze it all at once.
Solution Approach 2:
The patent extracts and removes the most problematic high abundance proteins from the blood sample before gel analysis. By using affinity columns or other separation methods to pre-clear the sample, the technique reduces the effective complexity of the proteome being analyzed, making it more manageable and enabling identification of diagnostic markers.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for early detection of diseases by identifying unique molecular fingerprints in blood, facilitating the diagnosis of conditions such as cancer and other biological disorders with improved reproducibility and sensitivity, enabling personalized medicine and predictive diagnostics.
Implementation Method 1
using detection reagents like antibodies or aptamers to measure protein levels in blood samples
Implementation Method 2
using detection reagents like antibodies or aptamers to measure protein levels in blood samples
Data Source
AI summary
The present invention relates generally to methods for identifying organ-specific secreted proteins and for identifying organ-specific molecular blood fingerprints therefrom. As such, the present invention provides compositions comprising such proteins, detection reagents for detecting such proteins, and panels, and arrays for determining organ-specific molecular blood fingerprints.